title: Kinetic model for signal binding to the Quorum sensing regulator LasR.
abstract: We propose a kinetic model for the activation of the las regulon in the opportunistic pathogen Pseudomonas aeruginosa. The model is based on in vitro data and accounts for the LasR dimerization and consecutive activation by binding of two OdDHL signal molecules. Experimentally, the production of the active LasR quorum-sensing regulator was studied in an Escherichia coli background as a function of signal molecule concentration. The functional activity of the regulator was monitored via a GFP reporter fusion to lasB expressed from the native lasB promoter. The new data shows that the active form of the LasR dimer binds two signal molecules cooperatively and that the timescale for reaching saturation is independent of the signal molecule concentration. This favors a picture where the dimerized regulator is protected against proteases and remains protected as it is activated through binding of two successive signal molecules. In absence of signal molecules, the dimerized regulator can dissociate and degrade through proteolytic turnover of the monomer. This resolves the apparent contradiction between our data and recent reports that the fully protected dimer is able to "degrade" when the induction of LasR ceases.
authors.affiliation: Biomedical Engineering, Department of Electrical Engineering, Ørsteds Plads 349, Technical University of Denmark, Kongens Lyngby DK-2800, Denmark. tsams@dtu.dk.
authors.forename: Anetta
authors.initials: A
authors.lastname: Claussen
authors.forename: Tim Holm
authors.initials: TH
authors.lastname: Jakobsen
authors.forename: Thomas
authors.initials: T
authors.lastname: Bjarnsholt
authors.forename: Michael
authors.initials: M
authors.lastname: Givskov
authors.forename: Martin
authors.initials: M
authors.lastname: Welch
authors.forename: Jesper
authors.initials: J
authors.lastname: Ferkinghoff-Borg
authors.forename: Thomas
authors.initials: T
authors.lastname: Sams
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008954:Models, Biological; D055503:Protein Multimerization / Q000502:physiology*; D059748:Proteolysis; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000378:metabolism*; D053038:Quorum Sensing / Q000502:physiology*; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; C080365:LasR protein, Pseudomonas aeruginosa; D015534:Trans-Activators